| Added by | pmartino |
|---|---|
| Group name | EquipePM |
| Item Type | Journal Article |
| Title | Solubility Characterization and Imaging of Intrabodies Using GFP-Fusions |
| Creator | Renaud et al. |
| Author | Emilie Renaud |
| Author | Pierre Martineau |
| Author | Laurence Guglielmi |
| Abstract | Ectopically expressed intracellular recombinant antibodies, or intrabodies, are powerful tools to visualize proteins and study their function in fixed or living cells. However, many intrabodies are insoluble and aggregate in the reducing environment of the cytosol. To solve this problem, we describe an approach based on GFP-tagged intrabodies. In this protocol, the GFP is used both as a folding-reporter to select correctly folded intrabodies and as a fluorescent tag to localize the scFv and its associated antigen in eukaryotic cells. Starting from a scFv gene cloned in a retroviral vector, we describe retrovirus production, cell line transduction, and soluble intrabody characterization by microscopy and FACS analysis. |
| Publication | Methods in Molecular Biology (Clifton, N.J.) |
| Volume | 1575 |
| Pages | 165-174 |
| Date | 2017 |
| Journal Abbr | Methods Mol. Biol. |
| Language | eng |
| DOI | 10.1007/978-1-4939-6857-2_9 |
| ISSN | 1940-6029 |
| Library Catalog | PubMed |
| Extra | PMID: 28255879 |
| Tags | Aggregation, Cell Separation, corresponding, Degradation, first, Flow Cytometry, Folding, Genes, Reporter, GFP, Green Fluorescent Proteins, HeLa Cells, Humans, Intrabody, last, MCF-7 Cells, Microscopy, original, Protein Engineering, Recombinant Fusion Proteins, Retroviridae, scFv, Single-Chain Antibodies, Solubility, Transduction, Genetic |
| Date Added | 2019/01/17 - 09:54:48 |
| Date Modified | 2021/03/05 - 10:46:45 |
| Notes and Attachments | PubMed entry (Attachment) |
Institut de Recherche en Cancérologie de
